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P.O.

Box 2345, Beijing 100023,China World J Gastroenterol 2004;10(1):122-126


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E-mail: wjg@wjgnet.com www.wjgnet.com Copyright © 2004 by The WJG Press ISSN 1007-9327

• CLINICAL RESEARCH •
Effects of exercise on lipid metabolism and musculoskeletal
fitness in female athletes
Kung-Tung Chen, Rong-Sen Yang

Kung-Tung Chen, Department of Genenal Education, Ming Hsin (186.44±24.90 mg/dL vs 193.00±23.48 mg/dL) were higher,
University of Science and Technology, Hsinchu 304, Taiwan, China but the HDL was significantly lower (62.18±10.68 mg/dL
Rong-Sen Yang, Department of Orthopaedics, College of Medicine, vs 52.26±4.84 mg/dL, P<0.05) in postmenopausal athletes
National Taiwan University, Taipei 10043, Taiwan, China (5/36) group than in postmenopausal control group (9/30).
Supported by the National Science Council of Taiwan, NSC91-2413-
H-159-001
Correspondence to: Dr. Rong-Sen Yang, Department of Orthopaedics, CONCLUSION: Postmenopausal athletes (5/36) who no
National Taiwan University Hospital, No.7 Chung-Shan South Road, longer took competing exercises had reduced levels of
Taipei 10043, Taiwan, China. yang@ha.mc.ntu.edu.tw physical activity, faced increased risk of cardiovascular
Telephone: +886-2-2312-3456 Ext 3958 Fax: +886-2-23936577 disease compared to active athletes (31/36) and the
Received: 2003-10-16 Accepted: 2003-11-20 postmenopausal controls (9/30). We may thus concluded
that long term exercise effectively improves musculoskeletal
fitness and prevents BMD loss in female athletes.
Abstract
Chen KT, Yang RS. Effects of exercise on lipid metabolism and
AIM: This study investigated the effects of intense training musculoskeletal fitness in female athletes. World J Gastroenterol
on lipid metabolism, bone metabolism and bone mineral 2004; 10(1): 122-126
density (BMD) in female athletes. http://www.wjgnet.com/1007-9327/10/122.asp

METHODS: Sixty-six female subjects participated in this study,


age ranging from 18 to 55 years. The sample group included
thirty-six athletic subjects and the control group comprised INTRODUCTION
thirty non-athletic individuals. Five athletes competed with Weightlessness or immobilization, as experienced by astronauts
national level (5/36) and nine non-athletic subjects (9/30) were in space, is a well known cause of significant and rapid bone
postmenopausal women. The assessment items included body mineral loss[1-24]. Furthermore sedentary individuals generally
composition, radius BMD, calcaneus BMD, lung function, have a lower bone mass than physically active individuals,
muscular endurance, renal and liver function, bone marker moderate exercise is known to increase skeletal mass[3]. The
assay and hormone status. All data were analysed, using SPSS above effect is most obvious in sports that place a significant
10.0 software, and were presented as mean rank statistical stress on the skeleton. Investigations of athletes have identified
difference, using the Kurskal-Wallis (K-W) test. After that the physical activity as a major determinant of bone mass in the
non-parameter statistics were used. Either K value or P value general population.
below 0.05 was considered significant. Physical fitness significantly influences quality of life. In
Taiwan, medical care quality and public health environment
RESULTS: Urine deoxypyridinoline/creatinine (Dpd/Cre) levels have improved markedly over recent decades. The incidence
increased significantly (5.93±2.31 vs 6.85±1.43, K<0.01), of fatal infectious diseases thus has reduced significantly and
sit-reach (29.30±9.48 cm vs 41.31±9.43 cm, K<0.001, the life span of Taiwanese has enlongated. Simultaneously,
P<0.001), 1 minute sit-ups with bended knees (1 min sit- the incidence of chronic diseases has increased, yet people
ups) (17.60±9.34 count vs 30.00±10.38 count, K<0.001, remain ignorant of the importance of exercise[1-5].
P<0.001), and vertical jump (25.27±6.63 cm vs 34.69±7.99 Exercising for 20-60 min per day, three days per week, at
cm, K<0.001, P<0.001) improved significantly in the athletes moderate intensity level of 3-6 Metabolic Equivalent units (METs)
group. The athletes group also had a significantly increased for most individuals derives at least some health-related benefits,
level of estriol (E3) (0.14±0.13 pg/mL vs 0.07±0.04 pg/mL, including improved cardiorespiratory fitness, muscle strength and
K<0.01, P<0.01), radius BMD (1.37±0.49 gm/cm2 vs 1.19±0.40 endurance, flexibility and body composition, as well as associated
gm/cm2, K<0.05) and calcaneus BMD (0.57±0.17 gm/cm2 psychological benefits. Consequently, lifelong physical exercise
vs -0.20±0.17 gm/cm2, K<0.01, P<0.05) compared with is recommended to optimize health-related benefits[2-8]. And
those of the controls. The high density lipoprotein (HDL) the influence of physical activity and exercise training on BMD
(65.00±14.02 mg/dL vs 52.26±4.84 mg/dL, K<0.05, P<0.05) in females previously has been assessed in cross-sectional,
was significantly lower in postmenopausal inactive athletes retrospective longitudinal and controlled trial studies[3-13].
(5/36) than premenopausal active athletes (31/36). On the Even though no relationship about growth hormone and
other hand, low-density lipoprotein (LDL) (98.35±23.84 mg/dL BMD was found. But the effect of E3 significantly improved
vs 131.00±21.63 mg/dL, K<0.05, P<0.01), cholesterol (CHO) BMD by inhibiting bone resorption, female athletes with low
(164.03±27.01 mg/dL vs 193.00±23.48 mg/dL, K<0.05, estradiol (E2) level take a risk for increased lipid peroxidation
P<0.05), triglyceride (TG) (63.00±26.39 mg/dL vs 147.00± following exercise[11-15]. Thus hormone status and lipid metabolism
87.21 mg/dL, K<0.01), body fat % (BF%) (28.16±4.90% vs may play an important role in the protection against cardiovascular
34.84±4.44%, K<0.05, P<0.001) and body mass index (BMI) disease, this physiological response has implications for risks of
(21.98±2.98 kg/m2 vs 26.42±5.01 kg/m2, K<0.05, P<0.001) heart disease. Longitudinal information on associations between
were significantly higher in postmenopausal inactive athletes life style factors and age-related bone loss remains quite
(5/36) than premenopausal active athletes (31/36). TG controversial. Some studies have found no relationship between
(90.22±39.82 mg/dL vs 147.00±87.21 mg/dL), CHO bone loss and body composition or body weight, while others
Chen KT et al. Lipid metabolism and musculoskeletal fitness in female athletes 123

have shown them to predict bone mass changes[3-16]. measured using an EIA kit obtained from Metra Biosystems
Therefore, the purpose of this study was to explore the (Monutain View, CA, USA). Urine Dpd level was measured
physiological function of female athletes, including BMD, renal using enzyme immunoassay (Ciba-Corning ACS-180) kits
function, liver function, hormone status, bone marker assay, purchased from Bayer international (Bayer Diagnostics,
lipid metabolism and muscle biology related to the effectiveness Tarrytown, NY, USA).
of exercise intervention for the health status of female athletes
compared with controls. BMD determination
Calcaneus site BMD was measured via speed of sound (SOS)
MATERIALS AND METHODS equipped for a bone mineral densitometry (Aloka Medical Ltd,
modelAOS-100, Tokyo, Japan) and all BMD values were also
Subjects expressed as a T-score, accurately reflecting the BMD. Distal site
Sixty-six female subjects participated in this investigation, with BMD was measured using the osmometer DTX-100 (SPA, Single
ages ranging between 18 and 55 yrs. The sample group was Photon Absorptiometry, Osmometer, Rodovre, Denmark). The
the athlete group (n=36), while the control group comprised scanners were calibrated daily against the standard calibration
non-athletic individuals (n=30). Inclusion criteria were that block supplied by the manufacturer to control baseline drift.
the female athletes had participated in high-intensity resistance
or impact activities (e.g., basketball, dancing). Exclusion Statistical analysis
criteria for both the subjects and the controls were that the All data were analysed, using SPSS 10.0 software, and were
subjects had no major medical illnesses, including coronary presented as mean rank statistical difference, using the Kurskal-
artery disease which could influence lipid metabolism, and Wallis (K-W) test. After that the non-parameter statistics were
were free of other risk factors that are associated with be used. The confidence interval was set at 95% and the
influencing lipid metabolism, such as smoking or ethanol intake significance level used was K<0.05 (two sides). All statistical
or treatment within the last two years with systemic gluco- analyses were carried out with SPSS statistical package. The
mineralocorticoids, anticonvulsants, bisphosphonates, Kruskal-Wallis test does not use any information on the relative
oestrogen, or raloxifene. Five athletes competed with national magnitude of each observation when compared with every other
level (5/36) and nine non-athletic subjects (9/30) were included observation in the combined sample. This comparison is replaced
in the analysis of postmenopausal women. The parameters to in each observation by its rank in the pool sample. The smallest
be measured included body composition, radius BMD and observation is replaced by its rank 1, the next smallest by rank
calcaneus BMD, lung function, muscular endurance, renal 2, and so on, the largest by its rank n. Since the test is an
function, liver function and hormone status. extension of the Mann-Whitney-Wilcoxon (M-W-W) test.
Either K value or P value below 0.05 is considered significant.
Anthropometric measurement of body composition
Anthropometric measurements were taken based on conventional
criteria. The measurement procedures of body weight (Wt) RESULTS
and body height (Ht) were estimated to the nearest 0.1 kg and No difference in body composition
0.5 cm, respectively. Finally BMI was calculated using the The thirty-six female athletes enrolled in this cross- sectional
formula: BMI (kg/m2)=Wt (kg)/Ht (m2). study did not differ significantly in terms of BF, BF%, BMI
and resistance compared with the control group (Table 1).
Health related fitness
They were tested using a modified Guthrie R test[6]. Health Table 1 Body composition of two groups
related fitness tests included vertical jump, 3 min steps, sit-
reach, hand grip and 1 min sit-ups items. 66 females

Variables Control group Athlete group K-Value


Lung function n=30 n=36
Respiratory muscle strength and pulmonary function were mean rank
assessed by spirometry. The flow volume and respiratory
muscle forces were measured using a Fukuda, microspiro HI- Body fat 31.68 35.01 0.483
501 model spirometer. BF% 33.83 33.22 0.898
BMI 32.45 34.38 0.685
Renal and liver function Resistance 34.95 32.29 0.575
Sixty-six blood samples per subject were drawn from an
antecubital vein with the subjects in the seated position. Routine Exercise improvements muscular endurance in female athletes
complete blood counts (CBC) were taken using a Sysmex-E9000 These two different groups did not differ significantly in muscular
(TOA Electronic, Inc., Tokyo, Japan) and renal and liver function endurance. The hand grip (28.06±6.14 kg vs 26.85±5.73 kg),
tests were performed using a Hitachi 7170 instrument (Hitachi 3 min steps (55.32±6.90 count/min vs 57.82±7.21 count/min) and
Electronic, Inc., Tokyo, Japan) by clinical chemistry laboratory vital capacity (86.86±15.98 L vs 88.23±12.05 L) in athlete group
staff at Li-Shin Hospital, Taoyuan County, Taiwan. were better than those in control group, but did differ significantly
in terms of sit- reach (29.3±9.48 cm vs 41.31±9.43 cm, K<0.001,
Hormone status P<0.001), 1 min sit-ups (17.60±9.34 count vs 30.00±10.38 count,
E2, E3, triiodothyronine (T3), thyroxine (T4), thyroid stimulating K<0.001, P<0.001) and vertical jump (25.27±6.63 cm vs
hormone (TSH), parathyroid hormone (PTH), cortisol and 34.69±7.99 cm, K<0.001, P<0.001) as listed in Table 2.
human growth hormone (HGH) were assayed in basal
conditions, using commercial radioimmunoassay (RIA) and Lipid metabolism
enzymeimmunoassay (EIA) Kits. Table 3 shows that the results were not significantly different
between both groups. But lipid metabolism including HDL
Bone marker assay (59.36±12.23 mg/dL vs 63.23±13.83 mg/dL) and Hb (12.95±1.22
Serum bone specific alkaline phosphatase (BAP) activity was g/dL vs 13.43±1.09 g/dL) in the athlete group was higher than
124 ISSN 1007-9327 CN 14-1219/ R World J Gastroenterol January 1, 2004 Volume 10 Number 1

in the control group. However, LDL (105.93±30.76 mg/dL vs Table 4 Serum enzyme activities related to renal and liver
102.89±25.92 mg/dL), TG (81.53±49.53 mg/dL vs 74.60±48.31 metabolism
mg/dL) and CHO (170.20±32.20 mg/dL vs 168.06±28.13 mg/dL)
were lower in the athlete group than those of the control group. Group n ALP Cre ALB DBIL
Thus exercise could improve the lipid metabolism, and it is
good for health. Control group (mean rank) 30 27.07 27.88 27.77 27.65
Athlete group (mean rank) 36 38.86ac 38.18ab 38.28a 38.38a
Table 3 No significant differences in blood CHO and lipid
variables between both groups a
K<0.05 vs statistically significant when compared with con-
trol group. bP<0.05 vs statistically significant when compared
Group n HDL LDL CHO TG Hb with control group. cP<0.01 vs statistically significant when
compared with control group.
Control group 30 31.92 34.65 34.92 35.02 28.75
(mean rank)
Athlete group 36 34.82 32.54 32.32 32.24 37.46
Renal and liver function
Table 4 shows that no difference between the data (data not
(mean rank)
shown here) of the two groups in terms of blood enzymes such

Table 2 Muscular strength and endurance assessment among controls and athlete groups

Group n Sit-reach 1 min sit-ups Vertical jump Hand grip 3 min steps Vital capacity

Control group (mean rank) 30 22.18 22.05 22.23 31.27 29.60 30.58
Athlete group (mean rank) 36 42.93ab 43.04ab 42.89ab 35.36 36.75 35.93

a
K<0.001 vs statistically significant when compared with control group. bP<0.001 vs statistically significant when compared with
control group.

Table 5 BMD, urine electrolytes, blood electrolytes in two groups

Group n Urine-Cre Blood-Ca BMD/radius BMD /calcaneus Blood-Cl

Control group (mean rank) 30 26.87 42.42 28.38 26.23 45.90


Athletes group (mean rank) 36 39.03ad 26.07b 37.76a 39.56bd 23.17c

a
K<0.05 vs statistically significant when compared with control group. bK<0.01 vs statistically significant when compared with
control group. cK<0.001 vs statistically significant when compared with control group. dP<0.05 vs statistically significant when
compared with control group.

Table 6 Hormonal findings in athletes with significance by non-parameter statistics test compared with controls

Group n Cortisol E3 T3 T4 PTH HGH

Control group (mean rank) 30 32.28 25.78 31.77 36.43 31.37 35.65
Athletes group (mean rank) 36 34.51 39.93ab 34.94 31.06 35.28 31.71

a
K<0.01 vs statistically significant when compared with control group. bP<0.01 vs statistically significant when compared with
control group.

Table 7 Biochemical bone turnover markers and BMD in athletes with significance by non-parameter statistics test as compared
with controls

Group n Dpd Urine-Cre (24 hrs) Dpd/Cre BAP BMD/radius BMD/Calcaneus

Control group (mean rank) 30 25.87 27.23 25.72 20.10 28.38 26.23
Athletes group (mean rank) 36 39.86be 38.72ad 39.99b 43.83cf 37.76a 39.56b

a
K<0.05 vs statistically significant when compared with control group. bK<0.01 vs statistically significant when compared with
control group. cK<0.001 vs statistically significant when compared with control group. dP<0.05 vs statistically significant when
compared with control group. eP<0.01 vs statistically significant when compared with control group. fP<0.001 vs statistically
significant when compared with control group.

Table 8 Postmenopausal female athlete lipid metabolism compared to premenopausal active athletes

Athletes group n BF% BMI HDL LDL CHO TG Hb

Premenopausal (mean rank) 31 16.84 17.08 20.15 16.74 16.98 16.55 18.79
Postmenopausal (mean rank) 5 28.60ac 27.30be 8.30ac 29.40ad 27.90ac 30.60b 16.70

a
K<0.05 vs statistically significant when compared with premenopausal group. bK<0.01 vs statistically significant when com-
pared with premenopausal group. cP<0.05 vs statistically significant when compared with premenopausal group. dP<0.01 vs
statistically significant when compared with premenopausal group. eP<0.001 vs statistically significant when compared with
premenopausal group.
Chen KT et al. Lipid metabolism and musculoskeletal fitness in female athletes 125

as glutamic oxalocetic transminase (GOT), glutamic pyruvic Lipid metabolism in postmenopausal females
transminase (GPT), blood urea nitrogen (BUN), uric acid (UA), Results from this study show higher levels of TG (90.22±39.82
total protein (TP), globulin (GLO) and bilirubin (BIL). But mg/dL vs 147.00±87.21 mg/dL), CHO (186.44±24.90 mg/dL
the control group displayed significantly lower alkaline vs 193.00±23.48 mg/dL), but lower levels of HDL (62.18±10.68
phosphatase (ALP) (61.03±13.99 U/L vs 70.81±15.23 U/L, mg/dL vs 52.26±4.84 mg/dL, P<0.05), Hb (13.82±0.88 g/dL
K<0.05, P<0.01), ALB (4.52±0.18 g/dL vs 4.62±0.27 g/dL, vs 13.52±0.21 g/dL) in postmenopausal athletes (5/36) group
K<0.05), Cre (0.75±0.09 mg/dL vs 0.81±0.10 mg/dL, P<0.05, compared with the postmenopausal control group (9/30). This
K<0.05) and direct bilirubin (DBIL) (0.25±1.11 mg/dL vs implies that the effect is a cardiovascular disease risk for
0.29±0.8 mg/dL, K<0.05) than the athlete group. postmenopausal retired female athletes (Table 9).

Electrolytes and BMD


DISCUSSION
According to non-parameter statistical tests, both the radius BMD
(1.37±0.49 gm/cm2 vs 1.19±0.40 gm/cm2, K<0.05) and calcaneus The data in this study were expressed as mean x±s. Statistical
BMD (0.57±0.17 gm/cm2 vs -0.20±0.17 gm/cm2, K<0.01, significance in the mean values was evaluated by the Student’s
P<0.05), increased significantly in the athlete group compared t test. But in our study, only sixty-six female subjects participated
with those of the control group. Moreover, the athlete group’s in this investigation. Therefore, we use K-W test to analyze
body electrolytes such as urine-Cre (132.22±72.30 mg/dL vs the results of all tests. The Kruskal-Wallis test does not use
166.83±62.52 mg/dL, K<0.05, P<0.05), blood calcium (Ca) any information on the relative magnitude of each observation
(8.76±0.32 mg/dL vs 8.43±0.37 mg/dL, K<0.01) and chloride when compared with every other observation in the combined
(Cl) (99.94±2.41 meq/L vs 102.83±1.97 meq/L, K<0.001) sample. This comparison is replaced in observation by its rank
significantly decreased compared to the control group. in the pool sample. The smallest observation is replaced by its
rank 1, the next smallest by rank 2, and so on, the largest by its
Hormone status rank n. Since the test is an extension of the M-W-W test. Either
K value or P value below 0.05 was considered significant.
HGH and T4 were lower in the athlete group than in the control
Exercise is important for maintaining skeletal health.
group (8.95±1.51 µg/dL vs 9.38±1.51 µg/dL), but cortisol
(11.39±4.03 µg/dL vs 10.75±3.42 µg/dL), E2 (88.82±66.42 pg/mL However, the ability of exercise to influence bone might not
be entirely related to hormone status. This study has shown
vs 80.56±63.10 pg/mL), T3 (112.07±13.52 ng/dL vs 114.78±17.16
that hormones and exercise interact to influence bone
ng/dL) and PTH (39.07±16.97 pg/mL vs 34.70±11.66 pg/mL)
levels were higher. Notably, E3 level (0.14±0.13 pg/mL vs adaptations, and thus raise E3 level related to increased BMD
following exercise in female athletes. For example, serum E2,
0.07±0.04 pg/mL, K<0.01, P<0.01) significantly increased in
cortisol, PTH and T3 levels in the athlete group were higher
the athlete group compared to those of the control group.
than those of the controls, and the major finding of this study
was that increased radius BMD (K<0.05) and calcaneus BMD
Bone marker assay and BMD (K<0.01, P<0.05) were significantly and positively related to
All biochemical and bone turnover markers, for example, serum E3 (K<0.01, P<0.01) concentrations[10-12]. Therefore, a
(67.97±39.67 nmol/mmol vs 102.63±46.97 nmol/mmol, clear understanding the interaction suggested by the present
K<0.01, P<0.01), Dpd/Cre ratio (5.93±2.31 vs 6.85±1.43, data between E3 concentration and the adaptation of bone to
K<0.01), and BAP (14.04±3.31 µg/L vs 20.93±6.17 µg/L, exercise is important, and provides an interaction through
K<0.001, P<0.001) significantly increased in the female which the estrogen receptors involved in the early response of
athlete group compared to those of the control group. The bone cells might increase their responsiveness to loading[11,12].
athletes displayed positive correlation of regional radius BMD These results indicate that physical exercise positively affects
(K<0.05) and calcaneus BMD (K<0.01, P<0.05) with these the maintenance of radius BMD (K<0.05), calcaneus BMD
results (Table 7). (K<0.01, P<0.05) in female athletes, thus increased E3 level
can prevent BMD loss and possible risk of osteoporosis[12].
Lipid metabolism in postmenopausal athletes The athletes have higher levels of all the biomarkers than the
Table 8 displays levels of LDL (98.35±23.84 mg/dL vs controls, including Dpd (K<0.01, P<0.01), urine-Cre (K<0.05,
131.00±21.63 mg/dL, K<0.05, P<0.01), CHO (164.03±27.01 P<0.05), Dpd/Cre ratio (K<0.005), BAP (K<0.001, P<0.001)
mg/dL vs 193.00±23.48 mg/dL, K<0.05, P<0.05), TG (63.00 and lower levels of blood-Ca (K<0.01), blood-Cl (K<0.001)
±26.39 mg/dL vs 147.00±87.21 mg/dL, K<0.01), BF% these results were associated with markedly increased radius
(28.16±4.90% vs 34.84±4.44%, K<0.05, P<0.001) and BMI BMD and calcaneus BMD[13-22].
(21.98±2.98 kg/m2 vs 26.42±5.01 kg/m2, K<0.05, P<0.001) Further studies are required to examine a larger population,
increased in the postmenopausal (5/36) inactive athletes group and also to consider the effects of BMD marker assay (for
compared to the premenopausal (31/36) active athletes. Then example insulin-like growth factors).
the level of HDL (65.00±14.02 mg/dL vs 52.26±4.84 mg/dL, Physical inactivity has been designated by the American
K<0.05, P<0.05) markedly decreased in the postmenopausal Heart Association as a major modifiable risk factor for
(5/36) inactive athletes. cardiovascular disease. Numerous studies have examined
individual morbidity and mortality from cardiovascular disease.
Table 9 Lipid metabolism of postmenopausal female athletes The results presented here indicate that exercise can improve
Postmenopausal n BMI HDL LDL CHO TG Hb
physiological characteristics, such the lowering levels of serum
group CHO and TG in female athletes, all of which may improve
cardiovascular fitness and reduce morbidity and mortality from
Control group 9 6.56 9.00 7.11 7.06 6.44 7.61 cardiovascular disease[12-16,23-26].
(mean rank) But the findings regarding the renal function, liver function
Athletes group 5 9.20 4.80a 8.20 8.30 9.40 7.30 and lipid metabolism of retired female athletes were surprising.
(mean rank) Enzyme activity indicates that this group (5/36) may not have
the same health benefits from physical exercise as the control
P<0.05 vs statistically significant when compared with post-
a subjects. Specifically, this group displayed decreased HDL
menopausal control group. (K<0.05, P<0.05), Hb and increased LDL (K<0.05, P<0.01),
126 ISSN 1007-9327 CN 14-1219/ R World J Gastroenterol January 1, 2004 Volume 10 Number 1

CHO (K<0.05, P<0.05), TG (K<0.01) compared to the 8 Hendriksen IJ, Meeuwsen T. The effect of intermittent training
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different lipoprotein phenotypes, the relations among total study in young Wistar rats. Bone 2002; 30: 293-299
CHO, LDL, HDL and CHD risk in older female athletes should 20 Hui SL, Perkins AJ, Zhou L, Longcope C, Econs MJ, Peacock M,
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ACKNOWLEDGEMENTS
21 Prestwood KM, Kenny AM, Kleppinger A, Kulldorff M.
We are grateful to Li-Shin Hospital in Taoyuan County, Taiwan Ultralow-dose micronized 17beta-estradiol and bone density and
that provided all laboratory tests in this study, which helped bone metabolism in older women: a randomized controlled trial.
us to complete the research subjects. JAMA 2003; 290: 1042-1048
22 van den Beld AW, de Jong FH, Grobbee DE, Pols HA, Lamberts
SW. Measures of bioavailable serum testosterone and estradiol
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